Note: Descriptions are shown in the official language in which they were submitted.
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" SYSTEMS AND METHODS FOR REDUCING LEUKOCYTES IN CELLULAR
PRODUCTS"
Field of the Invention
The invention generally relates to blood
processing systems and methods. In a more specific
sense, the invention relates to systems and methods
for removing leukocytes from blood components
collected for therapeutic purposes.
Background of the Invention
Today blood collection organizations
routinely separate whole blood into its various
therapeutic components, such as red blood cells,
platelets, and plasma.
One separation technique that is in
widespread use today uses a multiple blood bag
system. The bag system includes a primary blood bag
and one or more transfer bags, which are integrally
connected to the primary bag by tubing. The
technique collects from a donor a single unit (about
450 ml) of whole blood in the primary blood bag.
' The, donor is then free to leave.
The donor's whole blood later undergoes
centrifugal separation within the primary bag into
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red blood cells and plasma rich in platelets. The
plasma rich in platelets is expressed out of the
primary bag into a transfer bag, leaving the red
blood cells behind. The plasma rich in platelets
then undergoes further centrifugal separation within
the transfer bag into a concentration of platelets
and plasma poor in platelets. The plasma poor in
platelets is expressed from the transfer bag into
another transfer bag, leaving the concentration of
platelets behind.
Using multiple blood bag systems, all three
major components of whole blood can be collected for
therapeutic use. However, the yield for each
component collected is limited to the volume of the
components that are contained in a single unit of
whole blood. Furthermore, because red blood cells
are retained, United States governmental regulations
prohibit collecting another unit of whole blood from
the donor until six weeks later.
Certain therapies transfuse large volumes
of a single blood component. For example, some
patients undergoing chemotherapy require the
transfusion of large numbers of platelets on a
routine basis. Multiple blood bag systems simply
are not an efficient way to collect these large
numbers of platelets from individual donors.
On line blood separation systems are today
used to collect large numbers of platelets to meet
this demand. On line systems perform the separation
steps necessary to separate concentration of
platelets from whole blood in a sequential process
with the donor present. On line systems establish
a flow of whole blood from the donor, separate out
the desired platelets from the flow, and return the
remaining red blood cells and plasma to the donor,
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all in a sequential flow loop._
Large volumes of whole blood (for example,
2.0 liters) can be processed using an on line
system. Due to the large processing volumes, large
yields of concentrated platelets (for example, 4 x
1011 platelets suspended in 200 ml of fluid) can be
collected. Moreover, since the donor's red blood
cells are returned, the donor can donate whole blood
for on line processing much more frequently than
donors for processing in multiple blood bag systems.
Regardless of the separation technique
used, when collecting blood components for
transfusion, it is desirable to minimize the
presence of impurities or other materials that may
cause undesired side effects in the recipient. For
example, because of possible febrile reactions, it
is generally considered desirable to transfuse red
blood cells and platelets that are substantially
free of leukocytes, particularly for recipients who
undergo frequent transfusions.
Several United States Patents are directed
to the removal of leukocytes from red blood cells
and platelet components in multiple blood bag
systems. For example, see United States Patent
Number 4,767,541: 5,089,146; 5,100,564: and
5,128,048.
Nevertheless, a need still exists for
further improved systems and methods for removing
undesired matter like leukocytes from blood
components in a way that lends itself to use in high
volume, on line blood collection environments, where
higher yields of critically needed blood components
like platelets can be realized.
summary of the Invention
One aspect of the invention provides
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systems and methods for reducing the number of
leukocytes in a cellular rich concentrate using an
on line blood separation process.
As used in this Specification, the term "on
line blood separation process" refers to a blood
separation system or method that (i) establishes
communication between a source of blood and a flow
path: (ii) draws a blood volume from the source into
the flow path. and (iii) maintains communication
with the blood source for at least a portion of the
time that the blood volume undergoes separation
within the flow path.
As used in this Specification, an "on line
blood separation process" can separate the blood
volume either in a continuous manner or in an
interrupted manner. However, an "on line blood
separation process" maintains communication between
the flow path and the blood source for at least a
portion of the time the separation process occurs
within the flow path, regardless of specific timing
or sequencing of the separation process itself.
The on line blood processing system and
method provided by this aspect of the invention
includes a flow path that includes a device for
establishing communication between the path and a
source of blood. Separation means is associated
with the flow path for conveying blood from the
source into the flow path for a three stage
separation process.
The separation process serves in a first
stage to separate the blood within the flow path
into a cellular suspension.
The separation process serves in a second
stage to convey at least a portion of the cellular
suspension from the first separation stage for
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separation within the flow path that reduces the
number of leukocytes.
The separation process serves in a third
stage to convey the cellular suspension from the
second separation stage for separation within the
flow path that concentrates the cellular suspension
into a cellular-rich concentrate having a reduced
number of leukocytes.
The system and method include a control
mechanism that maintains communication between the
flow path and the blood source for at least a
portion of the separation process. It thereby
constitutes an on line blood separation process.
In a preferred embodiment, the control
mechanism maintains. communication between the flow
path~and the blood source substantially throughout
the separation process.
As used in this Specification, an "on line
blood separation process" can include external or
internal valves or clamps to interrupt flow within
the path to or from the blood source. However, in
the context of this Specification, such valves or
claims do not break the communication between the
blood source and the flow path. Instead, the valves
or clamps control fluid flow within the path while
maintaining communication between it and the blood
source.
Another aspect of the invention provides
systems and methods for reducing the number of
leukocytes in cellular concentrate :using an
substantially continuous blood separation process.
The system and method that embody this
aspect of the invention provide a three stage
separation process.
The process conveys blood for separation in
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a first stage into a cellular suspension.
The process conveys at least a portion of
the cellular suspension from the first stage for
separation in a second stage that reduces the number
of leukocytes. According to this aspect of the
invention, the conveyance into the second stage is
substantially continuous with separation occurring
in the first stage.
The process conveys the cellular suspension
from the second stage for separation in a third
stage that concentrates the cellular suspension into
a cellular-rich concentrate having a reduced number
of leukocytes. Also according to this aspect of the
invention, the conveyance into the third stage is
substantially continuous with separation occurring
in the second stage.
The invention is especially well suited for
the collection of platelet-rich concentrate having
a reduced number of leukocytes. The reduction of
the number of leukocytes from a platelet suspension
before its separation into a platelet concentration
does not activate platelets. It thereby leads to
effective and efficient leukocyte removal and a high
quality blood product suited for long term storage.
As used in this Specification, both the "on
line blood separation process" and the
"substantially continuous blood separation process"
differ from a multiple blood bag process. In a
multiple blood bag process, the blood source (i.e.,
the donor's circulatory system) does not~.remain in
communication with the flow path where separation of
the collected blood volume occurs. In a multiple
blood bag system, after a given blood volume is
collected in the primary bag, the donor's
circulatory system is disconnected from the primary
WO 95/03107 PCT/US94/01904
bag before separation occurs within the bag. Also,
in a multiple blood bag system, the separation
processes do not occur continuously. The first
stage separation of red blood cells and plasma rich
in platelets and the second stage separation of
platelets from plasma occur at different points in
time as separate, discontiguous steps.
Another aspect of the invention provides a
system for removing leukocytes from a cellular
suspension. The system includes two separation
devices. The first separation device separates
blood into a cellular suspension. The second
separation device reduces the number of leukocytes
in a cellular suspension.
According to this aspect of the invention,
the system includes a control mechanism that
operates the first and second separation devices in
a sequential process.
In this process, the system conveys blood
into the first separation device to undergo
separation into a cellular suspension. The system
also conveys a first portion of the cellular
suspension from the first separation device into the
second separation device to reduce the number of
leukocytes. At the same time, the system
recirculates a second portion of the cellular
suspension from the first separation device in a
path that bypasses the second separation device for
remixing with the blood that is being conveyed into
the first separation device.
By recirculating a portion of the cellular
suspension away from the second separation device,
the overall flow volume load on the second
separation device is reduced. This, in turn,
further enhances the leukocyte removal efficiencies
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of the system. When a platelet suspension is
recirculated, improved platelet collection efficiencies
are obtained in the first separation device.
The invention may be embodied in several forms
without departing from its spirit or essential
characteristics. The scope of the invention is defined
in the appended claims, rather than in the specific
description preceding them. All embodiments that fall
within the meaning and range of equivalency of the claims
are therefore intended to be embraced by the claims.
According to one aspect of the invention, there
is provided a system for obtaining a platelet suspension
having a reduced number of leukocytes comprising
a separation chamber having an entry region to
receive whole blood containing leukocytes to
centrifugally separate the whole blood into a first layer
comprising red blood cells, a second layer comprising a
platelet suspension which includes a number of
leukocytes, and an interface between the first and second
layers which includes a number of leukocytes, the
separation chamber including a collection region to
collect the platelet suspension and an outlet
communicating with the collection region,
an inlet path communicating with the entry
region of the separation chamber to convey whole blood
into the entry region,
an outlet path communicating with the outlet of
the separation chamber to convey platelet suspension from
the collection region,
a filter in the outlet path for removing
leukocytes from the platelet suspension,
a monitoring element monitoring location of the
interface within the collection region and transmitting
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location signals, and
a controller coupled to the monitoring element
and the outlet path and operating to continuously
maintain the location of the interface at a substantially
constant location in the collection region spaced from
the outlet, whereby the number of leukocytes in the
interface are retained in the separation chamber to
reduce the number of leukocytes present in the platelet
suspension, while platelet suspension is conveyed in the
outlet path from the collection region .through the
filter, whereby the number of leukocytes in the platelet
suspension is additionally reduced.
According to another aspect of the invention,
there is provided a system for obtaining a platelet-rich
concentrate having a reduced number of leukocytes
comprising
a separation chamber having a first portion to
receive whole blood containing leukocytes to
centrifugally separate the whole blood into a first layer
comprising red blood cells, a second layer comprising a
platelet suspension including a number of leukocytes, and
a third layer comprising a number of leukocytes between
the first and second layers, the separation chamber
including a second portion to separate the platelet
suspension into a platelet-rich concentrate and a
platelet-poor plasma constituent,
a whole blood path to convey whole blood into
the first portion of the separation chamber,
a platelet suspension path for conveying the
platelet suspension from the first portion to the second
portion of the separation chamber through an outlet, the
platelet suspension path including a filter which removes
leukocytes from the platelet suspension to provide a
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leukocyte-reduced platelet suspension for separation in
the second portion of the separation chamber, thereby
providing a platelet-rich concentrate having a reduced
number of leukocytes, and
a controller to continuously maintain the
location of the third layer at a substantially constant
location in the first portion of the separation chamber
spaced from the outlet, while separation of the
leukocyte-reduced platelet suspension occurs in the
second portion of the separation chamber, whereby the
number of platelets in the leukocyte-reduced platelet
suspension is additionally reduced before separation of
the leukocyte-reduced platelet suspension occurs in the
second portion of the separation chamber.
According to a further aspect of the invention,
there is provided a method for obtaining a platelet
suspension with reduced number of leukocytes comprising
the steps of
conveying whole blood containing leukocytes
into a centrifugal separation chamber for separation into
a first layer comprising red blood cells, a second layer
comprising a platelet suspension including a number of
leukocytes, and a third layer comprising a number of
leukocytes between the first and second layers,
at least partially while conveying whole blood
into the centrifugal separation chamber, conveying the
platelet suspension from the centrifugal separation
chamber through an outlet to a filter, while continuously
maintaining the third layer containing the number of
leukocytes at a substantially constant location inside
the centrifugal separation chamber spaced from the
outlet, whereby the number of leukocytes in the platelet
suspension is reduced, and
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filtering leukocytes from the platelet
suspension in the filter, whereby the number of
leukocytes in the platelet suspension is further reduced.
According to another aspect of the invention,
there is provided a method for obtaining a platelet
suspension with reduced number of leukocytes comprising
the steps of
conveying whole blood containing leukocytes
into a rotational field comprising a centrifugal
separation chamber for separation into a first layer
comprising red blood cells, a second layer comprising a
platelet suspension including a number of leukocytes, and
a third layer comprising a number of leukocytes between
the first and second layers,
conveying the platelet suspension from the
centrifugal separation chamber outside the rotational
field through an outlet to a filter, while continuously
maintaining the third layer containing the number of
leukocytes at a substantially constant location inside
the rotational field within the centrifugal separation
chamber spaced from the outlet, whereby the number of
leukocytes in the platelet suspension is reduced, and
filtering leukocytes from the platelet
suspension in the filter outside the rotational field,
whereby the number of leukocytes in the platelet
suspension is further reduced.
According to a further aspect of the invention,
there is provided a method for obtaining a platelet-rich
concentrate with reduced number of leukocytes comprising
the steps of
conveying whole blood containing leukocytes
into a centrifugal separation chamber for separation into
a first layer comprising red blood cells, a second layer
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comprising a platelet suspension including a number of
leukocytes, and a third layer comprising a number of
leukocytes between the first and second layers,
at Ieast partially while conveying whole blood
S into the centrifugal separation chamber, conveying the
platelet suspension from the centrifugal separation
chamber through an outlet to a filter, while continuously
maintaining the third layer containing the number of
leukocytes at a substantially constant location inside
the centrifugal separation chamber spaced from the
outlet, whereby the number of leukocytes in the platelet
suspension is reduced,
filtering leukocytes from the platelet
suspension in the filter, whereby the number of
leukocytes in the platelet suspension is further reduced,
and
conveying the leukocyte-reduced platelet
suspension from the filter to a second centrifugal
separation chamber for separation into a platelet-rich
concentrate and platelet-poor plasma.
According to another aspect of the invention,
there is provided a method for obtaining a platelet-rich
concentrate with reduced number of leukocytes comprising
the steps of
conveying whole blood containing leukocytes
into a rotational field comprising a centrifugal
separation chamber for separation into a first layer
comprising red blood cells, a second layer comprising a
platelet suspension including a number of leukocytes, and
a third layer comprising a number of leukocytes between
the first and second layers,
conveying the platelet suspension from the
centrifugal separation chamber outside the rotational
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field through an outlet to a filter, while continuously
maintaining the third layer containing the number of
leukocytes inside the rotational field at a substantially
constant location within the centrifugal separation
chamber spaced from the outlet, whereby the number of
leukocytes in the platelet suspension is reduced,
filtering leukocytes from the platelet
suspension in the filter outside the centrifugal field,
whereby the number of leukocytes in the platelet
suspension is further reduced, and
conveying the leukocyte-reduced platelet
suspension from the filter back into the rotational field
to a second centrifugal separation chamber for separation
into a platelet-rich concentrate and platelet-poor
plasma .
Brief Description of the Drawings
Fig. 1 is a diagrammatic view of a three stage
blood processing system that embodies the features of the
invention;
Fig. 2 is a plan side view of a blood
processing assembly that integrates two separation
elements of the system shown in Fig. 1;
Fig. 3 is a top view of the two element
assembly shown in Fig. 2;
Fig. 4 is a perspective view of the two element
assembly shown in Fig. 2, being partially wrapped upon a
centrifuge rotor for use and in association with the
remaining separation element of the system;
Fig. 5 is a side view, with portions broken
away and in section, of a centrifuge for rotating the
assembly shown in Fig. 4;
Fig. 6 is a partially diagrammatic view of a
multi-stage two needle blood processing system that
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incorporates the separation elements shown in Fig. 4;
Fig. 7 is a diagrammatic view of the two needle
blood processing system shown in Pict. 6; and
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Fig. 8 is a diagrammatic view of a process
for controlling the return and retention of
platelet-poor plasma in the system shown in Fig. 7.
Description of the Preferred Embodiments
Fig. 1 shows in diagrammatic form a
multiple stage blood processing system 10 that
embodies the features of the invention.
In use, the system 10 draws whole blood
(WB) from a donor, while adding anticoagulant. The
system 10 ultimately separates the anticoagulated WB
into three end products.
The first end product is red blood cells
(RBC). The second end product is a platelet-poor
component, which is commonly called platelet-poor
plasma (PPP). The third end product is a
resuspended, leukocyte-depleted platelet concentrate
(RES-LDPC).
The system 10 returns RBC to the donor.
The system 10 retains RES-LDPC for long term storage
and subsequent therapeutic use. The system 10
returns a portion of the PPP to the donor.
According to one aspect of the invention, the system
10 retains the rest of the PPP for various
processing purposes and for long term storage for
therapeutic purposes.
The system 10 employs three separation
stages to create these three end products.
In the first separation stage, the system
10 directs anticoagulated WB from the donor .into a
first separation element 12. The first.:element 12
separates the whole blood into RBC and an
intermediate product, which constitutes a platelet
suspension.
This platelet suspension is typically
plasma rich in platelets, and it is commonly
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referred to as platelet-rich plasma (PRP). However,
as used in this Specification, the term "platelet
suspension" is not limited to PRP in the technical
sense. The term "platelet suspension" is intended
to encompass any suspension in which platelets are
present in concentrations greater than in whole
blood, and can include suspensions that carry other
blood components in addition to platelets.
In the second separation stage, the system
10 directs PRP into a second separation element 14.
The second element 14 reduces the number of
leukocytes from the PRP, creating another
intermediate product, which constitutes leukocyte
depleted PRP (LDPRP).
As used in this Specification, the term
"leukocyte depleted" does not denote that all or
substantially all the leukocytes have been removed.
The term is intended to more broadly indicate only
that the number of leukocytes have been reduced by
some active separation process.
In the illustrated and preferred
embodiment, the system 10 diverts a portion of PRP
exiting the first element 12 away from the second
separation element 14. This-diverted flow of PRP is
recirculated back into the first separation element
12. The recirculated PRP joins the WB entering the
first separation element 12.
- In the third separation stage, the system
10 directs LDPRP into a third separation element 16.
The third element 16 separates LDPRP into another
intermediate product, which consists of leukocyte
depleted platelet concentrate (LDPC) and PPP.
As used in this Specification, the term
"platelet concentrate" is not limited to PC in the
technical sense. The term "platelet concentrate" is
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intended to encompass a volume of platelets that
results after a "platelet suspension" (as that term
is used in this Specification) undergoes a
subsequent separation step that reduces the fluid
volume of the platelet suspension.
In the illustrated and preferred
embodiment, the system 10 returns some of the PPP to
the donor during the processing period. The system
l0 retains the rest of the PPP during the processing
period.
The system 10 uses the retained PPP in
various transitional processing modes.
In one transitional mode, the system 10
adds retained PPP to LDPC. The PPP resuspends the
LDPC, creating the RES-LDPC end product. The PPP
serves as storage medium for the RES-LDPC during
long term storage. However, RES-LDPC can be used
for therapeutic use without long term storage.
In another transitional mode, the system 10
uses the retained PPP as an anticoagulated fluid to
rinse resident RBC from the system 10 after
processing for return to the donor.
In still another transitional mode, the
system 10 uses the retained PPP as an anticoagulated
fluid to keep fluid lines open and patent during
temporary interruptions in processing activity.
In still another transitional mode, the
system 10 recirculates the retained PPP to mix with
WB entering the first separation element l2. The
mixing of retained PPP with WB improves the
separation of RBC and PRP in the first element 12.
The system 10 collects the remaining
retained PPP for therapeutic purposes, with or
without long term storage.
EXAMPLE 1
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The exact constitution of the various blood
products created by the system 10 during process
varies according to the particular physiology of the
whole blood donor. Furthermore, as above indicated,
the terminology used in this Specification is not
intended to be restrictive.
Still, this Example is provided to describe
for purposes of illustration the constitution of the
blood products. that can be obtained from a healthy
donor.
A typical healthy donor has a WB hematocrit
(Hct) of between 40% (0.4) and 50% (0.5) before
donating WB. The hematocrit indicates the volume of
RBC per unit volume of WB. The healthy donor's WB
also contains about 250,000 platelets for each
milliliter (ml) of plasma.
After separation in the first separation
device 12, a portion of the plasma accompanies the
RBC that is returned to the donor. As a result, the
remaining platelet suspension (PRP) contains a
higher concentration of platelets than WB.
In PRP obtained from the typical donor,
there are about 350,000 to 400,000 platelets for
each ml of plasma.
The number of platelets per ml of PRP can
also be accurately estimated using the following
general expression:
P1a telets~
PlateletspRp = 1
CZ_Hct~
where PlateletsP~ is the concentration
(in number per ml) of platelets in the donor's PRP;
where PlateletsWB is the concentration
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(in number per ml) of platelets in the donor's WB
before donation. and
where Hct is the predonation
hematocrit of the donor's WB.
Essentially all the platelets carried in
PRP entering the third separate element 16 are there
separated from the plasma. The resulting LDPC takes
the consistency of a thick fluid-poor "paste" that
carries an extremely high concentration of platelets
l0 (approximately 30,000,000 to 40,000,000 platelets
per ml).
The LDPC is preferably resuspensed in about
200 ml of PPP to form RES-LDPC. The resulting RES
LDPC contains about 2,000,000 platelets per ml
resuspended in this fluid volume.
In the illustrated and preferred
embodiment, the system 10 comprises, once steril-
ized, a sterile, "closed" system, as judged by the
applicable standards in the United States.
Furthermore, the system 10 remains "closed" during
processing. This assures that the longest authorized
storage intervals can be used for the components
collected.
As will soon be apparent, the system 10
also lends itself to on line and/or continuous blood
separation processes.
The system 10 having the benefits of the
invention can be variously constructed.
In the illustrated and preferred
embodiments, the system 10 separates blood
components in both the first and third elements 12
and 16 using centrifugation techniques. Still, it
should be appreciated that other separation
techniques could be used for these purposes.
For example, the system 10 cari employ
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centrifugal separation techniques in conjunction
with the first element 12 and employ membrane
separation techniques in conjunction with the third
element 16. Such two stage processing techniques
using a combination of centrifugal and membrane
separation are disclosed in Schoendorfer U.S. Patent
4,851,126 and Kruger et al. U.S. Patent 4,680,025.
In the particular preferred embodiment
shown in Figs. 2 to 4, the system 10 integrates the
first and third elements 12 and 16 into a single
centrifugal processing chamber assembly 18.
Alternatively, the first and third elements 12 and
16 could comprise physically separate processing
chambers.
In the illustrated and preferred embodiment
(best seen in Fig. 4), a chamber assembly 18 is
formed of two sheets of flexible medical grade
plastic joined along their outer edge by a first
peripheral seal 20 (see Fig. 2, also).
A second interior seal 22 divides the
assembly 18 into a first processing compartment 24
and a second processing compartment 26. Although
part of an integral assembly 18, each processing
compartment 24 and 26 actually serves as a separate
and distinct separation element.
More particularly, the first compartment 24
comprises the first processing element 12. Here,
centrifugal forces separate whole blood into RBC and
PRP.
The second compartment 26 comprises the
third processing element 16. Here, centrifugal
forces separate LDPRP into LDPC and PPP.
Further details of the chamber assembly 18
are set forth in copending U.S. Patent Application
Serial No. 07/965,088, filed October 22, 1992 and
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entitled "Enhanced Yield Platelet Collection Systems
and Methods." This application is incorporated into
this Specification by reference.
In use, the chamber assembly 18 is wrapped
about a rotor 28 of a centrifuge 30 (see Figs. 4 and
5) .
The rotor 2 8 rotates about an axis 3 2 to
generate centrifugal forces. The centrifugal field
extends radially from the axis 32 through each
compartment 24 and 26. The compartment wall
radially spaced farther from the axis 32 will be
called the high-G wall 34 (see Figs. 3 and 4). The
compartment wall radially spaced closer to the axis
32 will be called the low-G wall 36.
The assembly 18 establishes a
circumferential flow of fluid during processing.
That is, the fluid introduced into each compartment
24 and 26 during rotation follows a circumferential
flow path in the compartment 24 and 26 about the
rotational axis 32.
In response to the centrifugal forces
generated in the first compartment 24, the higher
density RBC move toward the high-G wall 34,
displacing the lighter density PRP toward the low-G
wall 36 of the first compartment 24. An
intermediate layer called the interface forms
between the RBC and PRP. The interface constitutes
the transition between the formed cellular blood
components and the liquid plasma component.
Large amounts of leukocytes populate the
interface. When dynamic forces within the
compartment 24 are not optimized, platelets, too,
can settle out of the PRP and onto the interface.
In response to the centrifugal forces
generated in the second compartment 26, the higher
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d~ sity platelets in the PRP move toward the high-G
wall 34. They displace the lighter density liquid
PPP toward the low-G wall 36 of the second
compartment 26.
The construction and operation of the
centrifuge 30 can vary. Further details of the
centrifuge 30 are set forth in copending U.S. Patent
Application Serial No. 07/814,403, filed December
23, 1991 and entitled "Centrifuge with Separable
Bowl and Spool Elements Providing Access to the
Separation Chamber". This application is
incorporated into this Specification by reference.
Five ports 38/40/42/44/46 open into the
compartmentalized areas of the processing assembly
18. The ports 38/40/42/44/46 are arranged side-by
side along the top transverse edge of the respective
chamber 24 and 26. Three ports 38/40/42 serve the
first chamber 24. Two ports 44/46 serve the second
chamber 26.
The first port 38 comprises a PRP
collection port. The second port 40 comprises a WB
inlet port. The third port 42 comprises a RBC
collection port. The fourth port 44 constitutes a
PPP collection port. The fifth port 46 constitutes
a LDPRP inlet port.
The first chamber 24 includes a third
interior seal 48 (see Fig. 2) located between the
PRP collection port 38 and the WB inlet port 40.
The third seal 48 extends generally parallel to the
second interior seal 22 and then bends iri a dog-leg
away from the. WB inlet port 40. The dog-leg
terminates beneath the inlet of the PRP collection
port 38. The third interior seal 48 forms a PRP
collection region 50 within the first chamber 24.
The first chamber 24 also includes ~a fourth
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interior seal 52 located between the WB inlet port
40 and the RBC collection port 42. The fourth seal
52 extends generally parallel to the second and
third interior seals 22 and 48 and then bends in a
dog-leg away from the RBC collection port 42. The
dog leg terminates near the longitudinal side edge
of the first chamber 24 opposite to the longitudinal
side edge formed by the second interior seal 22.
Together, the third and forth interior
seals 48/52 form a WB inlet passage 54. Together,
the fourth interior seal 52, the second interior
seal 22, and the lower regions of the first
peripheral seal 20 form a RBC collection passage 56.
The WB inlet passage 54 channels WB
directly from the WB inlet port 40 into the flow
path at one end of the first chamber 24. WB enters
the circumferential flow path immediately next to
the PRP collection region 50. Here, the radial flow
rates of plasma are greatest, to lift platelets free
of the interface and into the PRP collection region
50.
The RBC collection passage 56 receives RBC
at the opposite end of the intended circumferential
flow path for WB within the chamber 24. From there,
the RBC collection passage 56 channels the RBC back
to the RBC collection port 42.
A ramp 58 (see Fig. 2) extends from the
high-G wall 34 of the~first compartment 24 across
the PRP collection region 50. The ramp 58 forms a
tapered wedge that restricts the flow. of fluid
toward the PRP collection port 38.
The ramp 58 also orients the interface
between RBC and PRP formed during separation for
viewing through a side wall of the chamber assembly
18 by an associated interface controller (not
WO 95103107 PCT/US94I01904
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18 -
shown). The interface controller monitors the
location of the interface on the ramp 58 and varies
the rate at which PRP is drawn from the chamber 24.
This holds the interface at a prescribed location on
the ramp 58, keeping RBC, white blood cells, and
lymphocytes away from the PRP collection port 38.
In the illustrated embodiment, a hinged
flap 60 (see Fig. 4) extends from and overhangs a
portion of the rotor. The flap 60 is preformed to
present the desired contour of the ramp 58.
Further details of interface control using
the ramp 130 are shown in Brown U.S. Patent
4,834,890, as well as in copending U.S. Patent
Application Serial No. 07/965,088, filed October 22,
1992 and entitled "Enhanced Yield Platelet
Collection Systems and Methods." Both are
incorporated into this Specification by reference.
The second compartment 26 includes a fifth
interior seal 62 (see Fig. 2) that extends between
the LDPRP inlet port 46 and the PPP collection port
44. The fifth seal 62 extends generally parallel to
the second seal 22 and then bends in a dog-leg away
from the LDPRP inlet port 46 in the direction of
circumferential PRP flow within the second chamber
26. The dog-leg terminates near the longitudinal
side edge of the second chamber 26 opposite to the
longitudinal side edge formed by the second interior
-seal 22.
The fifth interior seal 62, the second
interior seal 22, and the lower regions of: the first
peripheral seal 20 together form a PPP collection
passage 64. The PPP collection passage 64 receives
PPP at its open end and, from there, channels the
PPP to the PPP collection port 44.
In the illustrated and preferred embodiment
PCTlUS94101904
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(see Fig. 3), the low-G wall 36 of the first
compartment 24 is offset toward the high-G wall 34,
tapering into the compartment 24 in the direction of
circumferential WB flow. In the illustrated and
preferred embodiment (see Fig. 3), the low-G wall 36
also likewise tapers into the second compartment 26
in the direction of circumferential PRP flow.
In the illustrated and preferred embodiment
(see Fig. 2), the dog leg portion of the RBC
collection passage 56 and the dog leg portion of the
PPP collection passage 64 are both tapered in width
to present an enlarged cross section where they open
into their respective chamber 24 and 26. The
tapered widths of the passages 56 and 64 are each
preferably gauged, relative to the inward radial
taper of the low-G wall 36, to keep the cross
sectional area of the RBC collection passage 56 and
PPP collection passage 64 substantially constant.
The control of the cross section areas of
the collection passages 56 and 64 keeps fluid
resistance within the passages 56 and 64 relatively
constant. It also maximizes the available
separation and collection areas outside the passages
56 and 64. The tapered passages 56 and 64 also
facilitate the removal of air from the assembly 18
during pre-processing priming.
The tapering low-G wall 36 in the first
compartment 24 also preferably includes a stepped-up
barrier 66 (see Figs. 2 and 3) in the region where
the RBC collection passage 56 opens into the
compartment 24. The stepped-up barrier 66 extends
from the low-G wall 36 across the entire chamber 24,
as Fig. 2 shows. The stepped-up barrier 66 extends
into the RBC mass formed during separation, creating
a restricted passage 66 between it and the~facing
WO 95/03107 PCT/US94101904
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high-G wall 34 (see Fig. 3). The restricted passage
66 allows RBC present along the high-G wall 34 to
move beyond the barrier 66 for collection by the RBC
collection passage 56. Simultaneously, the stepped-
up barrier 66 blocks the passage of the PRP beyond
it, keeping the PRP within the dynamic flow
conditions leading to the PRP collection region 50.
Flexible plastic tubing attached to the
ports 38/40/42/44/46 interconnects the first and
second chambers 24 and 26 with each other, with the
second separation element 14, and with pumps and
other stationary components located outside the
rotating components of the centrifuge 30. The
flexible tubing is ganged together into an umbilicus
70 (see Figs. 4 to 6).
As Fig. 6 best shows, the non-rotating
elements attached to the umbilicus 70 include the
second separation element 14, the phlebotomy needles
78 and 80 that provide vein access to the donor, and
the various containers 80 that provide or receive
fluid during processing.
More particularly, the umbilicus 70
connects the WB inlet port 40 and the RBC collection
port 42 of the rotating assembly 18 with stationary
phlebotomy needles '78 and 80. One needle 78
continuously draws WB from the donor, while the
other needle 80 continuously returns RBC to the
donor. Alternatively, the system 10 could use a
single phlebotomy needle to perform both functions
in sequential draw and return cycles using
conventional techniques.
The umbilicus 70 also connects the PRP
collection port 38 of the first chamber 24 with the
LDPRP inlet port 46 of the second chamber 26, via
the second separation element 14, as Fig.~6 best
PCT/US94101904
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shows. The second chamber 26 thereby receives LDPRP
through the umbilicus 70 from the first chamber 24
(via the second separation element 14) for further
separation into PPP and LDPC. As Fig. 6 also shows,
a portion of the PRP exiting the PRP collection port
38 is diverted away from the second separation
element 14 for recirculation directly back to the WB
inlet port 40.
The umbilicus 70 also conveys separated PPP
from the second chamber 26 through the associated
PPP collection port 136. A portion of PPP is
conveyed by the umbilicus 70 to the donor return
needle 80. Another portion of PPP is conveyed by
the umbilicus to one or more of the collection
containers 81 for retention.
The LDPC remains behind in the second
chamber 26 for later resuspension and collection, as
will be described later.
As Fig. 5 shows, in operation, the
centrifuge 30 suspends the rotor 28 in an upside
down position during rotation, compared to the
position shown in Fig. 4.
As Fig. 5 also shows, a non-rotating (zero
omega) holder 72 holds the upper portion of the
umbilicus 70 in a non-rotating position above the
rotor 28.
Another holder 74 rotates the mid-portion
of the umbilicus 70 at a first (one omega) speed
about the rotor 28. Another holder 76 (see Fig. 4)
rotates the lower end of the umbilicus 70 next to
the assembly 18 at a second speed twice the one
omega speed (the two omega speed), at which the
rotor 28 also rotates. This known relative rotation
of the umbilicus 74 and rotor 28 keeps the umbilicus
74 untwisted, in this way avoiding the need for
WO 95/03107 PCT/US94/01904
c~ ~ ~:~:' - 2 2
rotating seals.
The dimensions of the various regions
created in the processing chamber can, of course,
vary according to the processing objectives. Table
1 shows the various dimensions of a representative
embodiment of a processing assembly 18 of the type
shown in Figs. 2 and 3. Dimensions A through F
referenced in Table 1 are identified in Figs. 2 and
3.
TABLE 1
Overall length (A): 19-1/2 inches
Overall height (B): 2-13/16 inches
First Stage Processing Chamber
Length (C): 10-1/8 inches
Width (D): 2-3/8 inches
Maximum Radial Depth in Use: 4 mm
Second Stage Processing Chamber
Length (E): 8-13/16 inches
Width (F): 2-3/8 inches
Maximum Radial Depth in Use: 4 mm
Port Spacing
(center line to center line) : 3/8 inch
In this configuration, the RBC collection
passage 56 and the PPP collection passage 64 taper
from a width of about 1/4 inch to 1/8 inch. The
restricted passage 68 in the first compartment 24 is
about 1 mm to 2 mm in radial depth and about 1 mm to
-2 mm in circumferential length.
In this configuration, when the rotor 28 is
rotated at a speed of about 3400 RPM, a centrifugal
force field of about 900 G's is generated along the
high-G wall 34 of the chambers 24 and 26.
Alternatively, the system 10 can employ
physically separate processing chambers as the first
and third elements 12 and 16. Such elements could
WO 95/03107 PCT/US94/01904
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then be usable in association with a commercially
available blood processing centrifuge, like the CS-
3000~ Blood Separation Centrifuge made and sold by
the Fenwal Division of Baxter Healthcare Corporation
(a wholly owned subsidiary of the assignee of the
present invention). These alternative processing
chambers are also disclosed in copending U.S. Patent
Application Serial No. 07/965,088, filed October 22,
1992 and entitled "Enhanced Yield Platelet
Collection Systems and Methods."
In the illustrated and preferred
embodiment, the system 10 carries out blood
component separation in the second element 14 using
filtration.
Still, the second element 14 can use other
separation techniques. The second element 14 can
separate leukocytes by centrifugation, absorption,
columns, chemical, electrical, and electromagnetic
means.
In the illustrated and preferred
embodiment, the second element 14 comprises a filter
device 82 that employs a non-woven, fibrous filter
media 84.
The composition of the filter media 84 can
vary. In the illustrated and preferred embodiment,
the media 84 comprises fibers that. contain nonionic
hydrophillic groups and nitrogen-containing basic
functional groups. Fibers of this type are
disclosed in Nishimura et al U.S. Patent 4,936,998,
which is incorporated herein by reference'. Filter
media 84 containing these fibers are commercially
sold by Asahi Medical Company in filters under the
tradename SEPACELL.
Filter media 84 containing these fibers
have demonstrated the capacity to remove leu~COCytes
WO 95/03107 PCTlUS94/01904
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while holding down the loss of platelets.
The system 10 shown in Figs. 1 to 6 can be
readily incorporated into a continuous single or
double needle on line blood processing systems.
Fig. 7 shows in diagrammatic form a representative
continuous two needle on line processing system 86
that carries out an automated resuspended platelet
collection procedure employing the features of the
invention.
l0 A processing controller 88 operates the two
needle system 86 in continuous collection and return
cycles, which occur simultaneously. As used in this
Specification, the terms "continuous" and
"simultaneous" are not meant to be limited to
sequences that are continuous or simultaneous only
in the quantitative sense. The terms are meant to
encompass sequences that, while not absolutely
continuous or simultaneous quantitatively, are
"substantially" continuous or simultaneous in a
qualitative sense, when there is no significant
operational or therapeutic difference in terms of
the way they operate and the function and result
achieved.
In the collection cycle, the donor's WB is
continuously supplied through the draw needle 78 to
the processing compartment 24 (for separation into
RBC and PRP), while continuously conveying PRP from
the compartment 24 into the filter device 82 (for
creating LDPRP), and while continuously conveying
LDPRP from the filter device 82 into the compartment
26 (for separation into LDPC and PPP),
During the return cycle, which in the two
needle system 86 occurs simultaneously with the
collection cycle, the controller 88 continuously
returns RBC from the compartment 24 and a portion of
WO 95/03107 PCTII1S94101904
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the PPP from the compartment 26 to the donor through
the return needle 80.
Throughout the processing period, the
circulatory system of the donor remains connected in
communication with the system 86 through the needles
78 and 80.
The controller 88 also continuously retains
a portion of the PPP exiting the compartment 26,
diverting it from return to the donor, during the
entire period that LDPC is being separated in the
compartment 26. The system 86 retains the.diverted
PPP for long term storage, as well as to aid
processing and resuspension of LDPC in the
compartment 26 for long term storage as RES-LDPC.
The system 86 includes a container 90 that
holds anticoagulant. While the type of
anticoagulant can vary, the illustrated embodiment
uses ACDA, which is a commonly used anticoagulant
for pheresis.
The system 86 also includes a container 92
that holds saline solution for use in priming and
purging air from the system 86 before processing
begins.
The system 86 also includes one or more a
collection containers 94 for receiving RES-LDPC for
therapeutic use, with or without long term storage.
The system 86 also includes one (or optionally more)
collection container 96 for retaining PPP during
processing. The container 96 ultimately can also
serve as a long term storage container for retained
PPP.
Under the control of the controller 88, a
first tubing branch 98 and a WB inlet pumping
station 100 direct WB from the draw needle 78 to the
WB inlet port 40 of the first stage processing
WO 95/03107 PCT/US94/01904
- 26
chamber 24. The WB inlet pumping station 100
operates continuously at, for example, 50 ml/min.
Meanwhile, an auxiliary tubing branch 102 meters
anticoagulant to the WB flow through an
anticoagulant pumping station 104.
ACDA anticoagulant can be added to
constitute about 9% of the entry WB. Saline
dilution fluid can also be added in an amount
representing about 4% of donor body volume (i.e.,
200 ml saline for 5000 ml in body volume).
Anticoagulated WB enters and fills the
first processing chamber 24 in the manner previously
described. There, centrifugal forces generated
during rotation of the chamber assembly 18 separate
WB into RBC and PRP.
The controller 88 operates a PRP pumping
station 106 to draw PRP from the PRP collection port
38 into a second tubing branch 108.
The processing controller 88 monitors the
location of the interface on the ramp 58. It varies
the speed of the PRP pumping station 106 to keep the
interface at a prescribed location on the ramp 58.
The controller 88 also limits the maximum rate of
the variable PRP pumping station 106 (for example,
25 ml/min) to be less than the WB inlet pumping
station 100.
Meanwhile, a third tubing branch 110
-conveys the RBC from the RBC collection port 42 of
the first stage processing chamber 24. The third
tubing branch 110 leads to the return needle 80 to
return RBC to the donor.
The system 86 includes a recirculation
tubing branch 112 and an associated recirculation
pumping station 114. The processing controller 88
operates the pumping station 114 to divert apportion
WO 95/03107 PCT/US94101904
_ 2~
of the PRP exiting the PRP collection port 38 of the
first processing compartment 24 for remixing with
the WB entering the WB inlet port 94 of the first
processing compartment 24.
The controller 88 can control the
recirculation of PRP in different ways.
In the illustrated and preferred
embodiment, the pumping rate of the recirculation
pump 114 is maintained as a percentage (%~) of the
pumping rate WB inlet pump 100 governed as follows:
%~ = K * Hct - 100
where:
Hct is the hematocrit of the donor's whole
blood, measured before donation, and
K is a dilution factor that takes into
account the volume of anticoagulant
and other dilution fluids (like
saline) that are added to the donor's
whole blood before separation.
When the pumping rate of the recirculation
pump 114 is maintained at the predetermined
percentage (%~) of the pumping rate WB inlet pump
100, a surface hematocrit of about 30% to 35% is
maintained in the WB entry region of the first
compartment 24. The preferred surface hematocrit in
the entry region is believed to be about 32%.
Keeping the surface hematocrit in the entry
region in the desired range provides maximal
separation of RBC and PRP.
The value of the dilution factor. K can vary
according to operating conditions. The inventor has
determined that K = 2.8, when ACDA anticoagulant is
added to constitute about 9% of the entry whole
blood volume, and a saline dilution fluid is added
in an amount representing about 4% of donor body
WO 95/03107 PCT/US94/01904
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28 -
volume (i.e., 200 ml saline for 5000 ml in body
volume) .
By mixing PRP with the WB entering the
first processing compartment 24 to control surface
hematocrit in the entry region, the velocity at
which RBC settle toward the high-G wall 66 in
response to centrifugal force increases. This, in
turn, increases the radial velocity at which plasma
is displaced through the interface toward the low-G
wall 64. The increased plasma velocities through
the interface elute platelets from the interface.
As a result, fewer platelets settle on the
interface.
The remainder of the PRP exiting the first
compartment 24 enters the filter device 82 (by
operation of the pumping station 106) for removal of
leukocytes. A preferred flow rate of .PRP through
the filter device 82 is in the range of 15 to 30
ml/m~nute.
The continuous on-line removal of
leukocytes from PRP that the invention provides does
not activate platelets carried in the PRP.
The concurrent recirculation of a portion
of the PRP away from the filter device 82 reduces
the overall flow volume load on the filter device
82. This, in turn, enhances the leukocyte removal
efficiencies of the filter device 82.
A fourth tubing branch 118 conveys LDPRP to
the LDPRP inlet port 46 of the second stage
processing chamber 26. There, LDPRP.:undergoes
further separation into LDPC and PPP, as earlier
described.
PPP exits the PPP collection port 44 of the
second stage processing chamber 26 through a fifth
tubing branch 120. The fifth tubing branch 120
WO 95103107 PCTlUS94/01904
29
joins the third tubing branch 110 (carrying RBC),
which leads to the return needle 80.
The system 86 includes a sixth tubing
branch 122 and an associated pumping station 124 for
continuously retaining a portion of the PPP during
the entire processing period.
The processing controller 88 operates the
PPP retaining pumping station 124 throughout the
period that separation occurs within the compartment
l0 26 to continuously divert a prescribed portion of
the PPP exiting the PPP collection port 44 away from
the third tubing branch 110. The sixth tubing
branch 122 continuously conveys the diverted PPP to
the collection container 96 for retention.
As before stated, the remainder of the PPP
exiting the second compartment 26 enters the third
branch 110, where it joins the RBC for return to the
donor by operation of the PPP return pump station
116.
The processing controller 88 controls the
flow rate of the pumping station 124 to retain the
desired proportional volume of PPP separated in the
second chamber 26. The system 86 returns the
remaining PPP, to the donor:
In the preferred embodiment, the controller
88 incorporates a PPP retention control process 132
(see Fig. 8) that governs the rate at which PPP
should be retained (expressed in ml/min) during the
processing period to collect the volume of PPP
desired by the end of the processing period. The
control process not only takes into account the
physical operating parameters of the system 86, but
it also takes into account the physiology and
comfort of the individual donor.
As Fig. 8 shows, the control process 132
WO 95/03107 PCT/US94/01904
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receives as input the donor's weight; the donor's
platelet count (before donation); the desired
platelet yield; and the targeted system efficiency.
The system efficiency is expressed as the percentage
of the platelets processed that are ultimately
collected as LDPC.
Taking these parameters into consideration,
the control process 132 determines the total WB
volume that should be processed to obtain the
desired platelet yield.
From the total WB volume, the control
process 132 derives the total PPP volume. The
plasma volume is derived by multiplying the total WB
volume by (1.0 - Hct), where Hct is expressed in
decimal form (e.g., 0.4 instead of 40%).
The control process 132 also receives as
input the amount of PPP the operator wants to retain
during processing. The control process 132 first
compares the desired PPP retention volume to the
maximum volume of PPP that, under applicable
governmental regulations, can be retained. In the
United States, for example, the Food and Drug
Administration limits the volume of plasma that can
be collected from a donor during~a given procedure
to 600 ml for donors with a body weight less than
175 pounds, and 750 ml from a donor with a body
weight equal to or greater than 175 pounds.
If the desired PPP retention volume does
not exceed the maximum allowable retention volume,
the control process 132 subtracts the desired PPP
volume from the total PPP volume to derive the PPP
return volume. Otherwise, the control process 132
generates an error signal, informing the user that
less PPP must be retained.
The control process 132 uses the donor's
WO 95/03107 PCT/US94/01904
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hematocrit (Hct) and the selected anticoagulant
ratio to set a WB flow rate the donor should be able
to tolerate. A look up table can be empirically
prepared to correlate WB flow rates with these
factors.
Alternatively, the user can select a
nominal WB flow rate based upon experience (say, 50
to 60 ml/min). The user can then make on line
adjustments as necessary during processing based
upon the observed response of the donor.
Based upon the WB processing volume and the
selected anticoagulant ratio, the control process
132 determines the volume of anticoagulant that will
be mixed with WB during processing.
The control process 132 adds together the
total WB processing volume and the total
anticoagulant volume (plus any dilution fluids, if
added) to determine the total fluid volume that will
be processed. By.dividing the total fluid volume by
the WB flow rate, the control process 132 calculates
the total processing time.
Based dividing the desired PPP retention
volume by the derived processing time, the control
process 132 derives the rate at which the PPP
retention pump 124 must be operated.
The control process 132 also determines the
effect of the PPP retention rate upon the rate at
which the donor receives citrate in the portion of
PPP that is returned to the donor. The control
process adjusts the derived PPP retention rate as
necessary to avoid a citrate return rate that is
higher than a preset, physiologically relevant
value.
Based upon the total volume of
anticoagulant and the type of the anticoagulant, the
WO 95103107 PCT/US94101904
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control process 132 determines the total volume of
citrate that will be mixed with WB.
The proportion of citrate contained in
anticoagulants is known. For example, ACDA
anticoagulant contains 21.4 mg of citrate for each
ml of solution.
Based upon the proportion of PPP that will
be returned to the donor, the control process 132
derives the rate at which the system will infuse the
citrate into the donor. The control process 132
derives the citrate infusion rate in terms of the
amount of citrate (in mg) the donor receives with
the returned PPP per unit of donor body weight (in
kg) per unit of processing time (in minutes).
The control process 132 compares the
derived citrate infusion rate to a nominal citrate
infusion rate, which is also expressed in mg
citrate per kg body weight per minute of processing
time. The nominal citrate infusion rate represents
a predetermined maximum rate that citrate can be
infused in a typical donor without causing donor
discomfort and other citrate-related reactions. The
control process 132 in the illustrated and preferred
embodiment sets the nominal citrate infusion rate at
1.2 mg/kg/min, which is based upon empirical data.
As long as the targeted PPP retention rate
results in a citrate infusion rate that is equal to
or less than 1.2 mg/kg/min, the controller 88 is
free to adjust the operating parameters of the
system 86 as necessary to maximize the~.-component
yields and processing efficiencies.
However, if a targeted PPP retention rate
results in a citrate infusion rate that exceeds 1.2
mh/kg/min, the controller 88 adjusts the processing
parameters to lower the citrate infusion rate.
WO 95103107 PCT/US94I01904
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The control process 132 shown in Fig. 8 can
be expressed in various ways in terms of a series of
simultaneous equations that, when solved, derive the
operating parameters listed above. The equations
can be solved recursively during processing to take
into account changes in Hct and platelet counts in
the donor's WB.
The continuous retention of PPP serves
multiple purposes, both during and after the
component separation process.
The retention of PPP serves a therapeutic
purpose during processing. PPP contains most of the
anticoagulant that is metered into WB during the
component separation process. By retaining a
portion of PPP instead of returning it all to the
donor, the overall volume of anticoagulant received
by the donor during processing is reduced. This
reduction is particularly significant when large
blood volumes are processed.
By continuously diverting PPP away from the
donor throughout the separation process, the donor
realizes this benefit continuously throughout the
processing period, provided that the nominal citrate
infusion rate is not exceeded. The monitoring
function that control process 132 performs assures
that this benefit will be obtained.
Also the retention of PPP during processing
keeps the donor's platelet count higher and more
uniform during processing. This is because the
donor receives back less fluid volume during the
processing period, thereby reducing the dilution of
whole blood undergoing processing.
EXAMPLE 2
This Example demonstrates the results
obtained by operation of the control process 132
WO 95/03107 PCT/US94/01904
_ 34 _
shown in Fig. 8 in connection with a platelet
collection procedure for a representative donor.
In this Example, the donor is assumed to
have a body weight of 125 lbs (56.3 kg). The donor
is also assumed to have a preprocessing Hct of 40%
and a preprocessing platelet count of 250,000 per
ml.
In this Example, the desired platelet yield
for the process is 4.0 x 1011; and the desired system
efficiency is 90%. ACDA anticoagulant is used at a
WB_to_anticoagulant ratio of 9%.
Based upon the above assumptions, the
control process derives 2.0 L as the total WB volume
that must be processed. The total volume of PPP
associated with this WB volume is 1200 ml.
Based upon the 9% anticoagulant ratio, the
control process 132 derives 180 ml as the volume of
anticoagulant that will be added. The total
processing volume is therefore 2180 ml. The total
citrate volume is 38.52 ml.
The user sets 60 ml/min as the nominal WB
inlet flow rate. At this rate, the processing time
will be 36.3 minutes.
The user seeks to ultimately retaining 500
ml of PPP during processing, which is well short of
the prescribed maximum for the donor s body weight.
With a processing time of 36.3 minutes, the PPP
-retention pump must be operated at a rate of 13.8
ml/min throughout the process to collect the desired
PPP volume of 500 ml.
The remaining 700 ml will be returned to
the donor. This means that the donor will receive
only about 58% (7/12'h) of the total anticoagulant
volume that would have been received, if the system
86 retained no PPP. '
WO 95103107 PCTIUS94/01904
~14383~
- 35 -
Moreover, when 500 ml of PPP is retained,
the total fluid volume returned to the donor is just
3/4 of the total drawn volume. In a single needle
system, each return cycle would take less time and
allow an overall higher blood flow rate.
The control process 132 derives the citrate
infusion rate by first determining the volume of
citrate that the returned PPP will carry, which is
22.47 ml (7/12'h of 38.52 ml). The control process
132 then divides the returned citrate volume by the
donor's body weight (56.3 kg) and by the processing
time (36.3 min) to obtain a citrate return rate of
1.1 mg/kg/min.
The control process 132 determines that the
citrate return rate is less than the nominal 1.2
mg/kg/min. The PPP retention pump 124 can thereby
be operated at 13.8 ml/min without anticipating
adverse donor citrate-related reactions.
The system 86 also derives processing
benefits from the retained PPP.
ALTERNATIVE RECIRCULATION MODE
. The system 86 can, in an alternative
recirculation mode, recirculate a portion of the
retained PPP, instead of PRP, for mixing with WB
entering the first compartment 24.
In this mode, the system 86 opens clamps C6
and C7 to convey retained PPP from the container 96
through a PPP recirculation branch 134 (see Fig. 7)
and associated pumping station 136 into the
recirculation branch 112. .:
The controller 88 controls the pumping
station 136 in the same manner described for pumping
station 114 to mix retained PPP with the incoming WB
entering the first compartment 24.
3 5 REEF-OPEN MODE
WO 95103107 PCTIUS94101904
_ 36 _
Should WB flow be temporarily halted during
processing, the system 86 enters a "keep-open" mode.
In this mode; the system 86 draws upon the retained
volume of PPP as an anticoagulated "keep-open"
fluid.
More particularly, the system controller 88
closes clamps C1 and C2, opens clamp C3, and
operates the pump 116 to direct a volume of the PPP
from the container 96 through a keep open tubing
branch 126, which communicates with the WB tubing
branch 98. The anticoagulant present in the PPP
keeps the WB tubing branch 98 open and patent until
the flow of anticoagulated WB resumes.
The system controller 88 keeps the return
needle 80 open and patent by intermittently closing
clamp C3 and opening clamp C2 to provide a keep open
flow of anticoagulated PPP through the return needle
80.
Use of PPP as a keep-open fluid avoids the
need to introduce additional anticoagulant from
container 90 for this purpose.
RINSE-BACR MODE
The system 86 also enters a "rinse-back"
mode at the end of the separation process. In this
mode, the system 86 draws upon the retained volume
of PPP as a "rinse-back" fluid.
More particularly, the system controller 88
stops the flow of WB through the tubing branch 98 by
closing clamp C4. The system controller 88 closes
the clamp C2, opens clamp C3, and operates the pump
116 to direct a volume of the collected PPP into the
first separation compartment 24 through tubing
branches 122, 120, 126 and 98. The flow of PPP
resuspends and purges RBC from the compartment 24
for return to the donor through the return' branch
WO 95/03107 PCTlUS94/01904
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110.
Because PPP, and not saline, is the rinse-
back fluid, downstream component separation through
the filter device 82 and the second compartment 26
can continue without interruption during the rinse
back mode. The PPP volume used for rinse-back
purposes ultimately returns, after separation in the
first compartment 24 and flow through the second
compartment 26, to the source PPP retention
container 96.
RESUSPENSION MODE
The system 86 also operates in a
resuspension mode to draw upon a portion of the
retained PPP to resuspend LDPC in the second
compartment 24 for transfer and storage as RES-LDPC
in the collection containers) 94.
In this mode, the system controller 88
closes clamps C2, C3, and C5, while opening clamp
C1. The controller 88 operates the pump 124 to
convey a volume of retained PPP through the PPP
collection port 44 into the second compartment 26.
This returned PPP volume mixes with and,resuspends
LDPC in the second compartment 26.
Preferably, during the resuspension period,
the compartment 26 is not rotated at high speeds.
Instead, as PPP is conveyed into the second
compartment 26, the rotor 28 slowly oscillates the
assembly 18 first in one rotational direction and
then in an opposite rotational direction. The
oscillation creates constantly changing acceleration
or agitation forces that aid the mixing and
resuspension process within the second compartment
26.
Also, a substantial portion of the fluid
volume residing with the LDPC in the' second
WO 95/03107 PCT/LJS94/01904
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38 -
compartment 26 is preferably drawn away before
introducing the resuspension volume of PPP. The
controller 88 operates the pump 124 to convey this
residual fluid volume from the second compartment 26
into the PPP retention container 96.
In the illustrated and preferred
embodiment, the second chamber 26 contains about 40
ml of PPP at the end of the processing period.
About 20 ml of the residual fluid volume is conveyed
away before resuspending the LDPC.
The second compartment 26 is therefore
about half empty of residual fluid when the
resuspension process begins. This fluid-depleted
condition concentrates the volume of LDPC to
intensify the acceleration forces generated when the
compartment 26 is oscillated. This further enhance
the mixing and resuspension~process.
In the illustrated and preferred
embodiment, about 20 ml of PPP is returned and used
as a resuspension fluid. After a predetermined
mixing period, the system controller 88 closes clamp
C1, opens, clamp C5, and operates the RES-LDPC
pumping station 128. This conveys the RES-LDPC
volume out the PPP collection port 44 through a RES-
LDPC collection tubing branch 130 into the
containers) 94 for storage.
In a preferred implementation, the
resuspension mode constitutes a series of sequential
resuspension batches. In each batch, the residual
fluid volume of the second compartment 26 is
depleted by about 20 ml, and the assembly 18 is
oscillated, and a prescribed aliquot of about 20 ml
PPP is conveyed into it.
Following a set mixing period, a volume of
LDPC that is resuspended is conveyed out ~of the
PCT/US94/01904
WO 95/03107
- '
second compartment 26 and into the containers) 94.
These sequential batches repeat, until all the
volume of LDPC is suspended and conveyed as RES-LDPC
into the collection containers) 94.
At the end of the resuspension process, a
resuspension volume of PPP (typically about 200 ml)
resides with the RES-LDPC to serve as a storage
medium. Typically, about 4.0 x 1011 platelets are
suspended in this fluid vo~,~me.
The remaining volume of retained PPP is
stored in the container 96 for subsequent
therapeutic use.
The system 86 thereby boosts the yield of
usable therapeutic components during a given
collection procedure.
In a single needle collection system (not
shown) that embodies the features of the invention,
a return cycle does not occur simultaneously with a
collection cycle. Instead, the single system
repeatedly toggles between a collection cycle and a
return cycle.
During a given collection cycle, WB is
drawn through the single needle, and the separated
RBC and a portion of -the separated PPP are
temporarily pooled in reservoirs. When a
preselected volume of RBC is pooled in this manner,
the associated controller switches to a return
cycle. In the return cycle, the WB flow from the
donor is suspended, while the pooled RBC and PPP are
returned to the donor through the same single
needle.
In a single needle system, processing can
continue uninterrupted through the compartment 24,
filter device 82, and the compartment 26 during a
return cycle by collecting a quantity of WB in a
WO 95!03107 PCT/US94101904
40 -
4r -
reservoir upstream of the compartment 24 during the
collection cycle. The single needle system then
draws WB from the reservoir for processing during a
return cycle.
A single needle system also retains a
portion of the PPP exiting the compartment 26,
diverting it from pooling and return to the donor,
during the entire period that LDPC is being
separated in the compartment 26. The retained PPP
is used for the same purposes in a single needle
system as in a two needle system.
The chamber assembly 18, the associated
containers, and the interconnecting tubing branches
associated with the system can be made from
conventional approved flexible medical grade plastic
materials, such .as polyvinyl chloride plasticized
with di-2-ethyl-hexylphthalate (DEHP).
Preferable, the containers) 94 intended to
store the resuspended LDPC, are made of polyolefin
material (as disclosed in Gajewski et al U.S. Patent
4,140,162) or a polyvinyl chloride material
plasticized with tri-2-ethylhexyl trimellitate
(TEHTM). These materials, when compared to DEHP-
plasticized polyvinyl chloride materials, have
' 25 greater gas permeability that is beneficial for
platelet storage.
Various features of the inventions are set
forth in the following claims.
